Download Haemoglobin, Transf errin and

Haemoglobin, Transf errin and
.Iburnin Types in Eqzdidae
Horses, Mules, Donkeys and
Zebras)
by
D. R. Osterhoff
Department of ZootechnoIogy,
Fnrulty of Ve'ierinary Science,
University of Pretoria, 0nderstepoor.l.
horse.;; the group consisted mainly of crossbrd
riding horses with rektive few family data available. These horses aIso provided the female basis
of the hybrids. A classification of the African donkey into breeds is almost impossible and no further
description is necessary.
The zebras belonged t o the species Eqerus burcheZl+
antiqztorum; most of the 38 samples included in the
present study originated from animals in the Etosha
Game Reserve, South West Africa. Several samples
wpre collected in the Kruger National Park and
other game reserves.
The determination of haernoglobins was performed
by using the method of starch gel electrophoresis as
described by Buschmann (1963) including the new
technique of the preparation of starch as originated
by ICristjansson 11963).
For transferrins the method of starch gel eIectrophorcsis as described by Kristjansson and Hickman
(1965)was applied in toto, the only difference being
the double slicing of the gel to facilitate check
readings of phenotypes.
Albumins were typed according to the method described by Braend and Efremov (1965) but also
here, the preparation of starch was done accordinz
to techniques described by Kristjansson 11963).
RESULTS
Fig. 1 shows the variation in haemoglobin obtained
in the genus equidae.
INTRODUCTION
Blood protein polymorphisrn provides the irnrnunogeneticist with interesting took for the search into
possible relationships between different species and
other problems of origin and domestication. In
equidae we are in the fortunate position to have at
hand three species of one genus and at the same
time a hybrid between two of these species. Several
Investigations of protein polymorphisrn (Bangham
and Lehmann, 1958; Podliachouk et al, 1965: Kaminski, 1965), enzyme differences (Kaminski and
Cajos, 1964),blood groups (Podliachouk, 1965 ; Podliachouk et al, 1965) and immunological differences
(Podliachouk et al, 1965) have been performed on
horses, donkeys and their hybrids, but so far no
zebns were included in any of these studies.
The present paper should be regarded as a preliminary study which will be folEowed by an investfgation on a greater zebra as well as donkey and
mule material with more family data incIuded i n
the latter.
METHODS AND MATERIAL
The material of 234 horses, 40 donkeys and 52
muIes formed a mixed group of animals which are
either bred a t o r purcP,ased for t h e Veterinary Research Institute, Onderstepoort. No clear classif ication of breeds can be given with regard to the
WNKEY
MULE
Figure 1. Haemoglobin types
t
DO
in cguidar.
Three haemoglobin phenotypes could be established
in horses, two of them being described before
(Braend and Effemov, 1965; Schmid, 1965) and
another phenotype which consisted also of two components but with a major band migrating slower
than a secondary faster moving band.
Using the nomenclature introducd by Braend and
Efremov (1965) and adding a plus o r minus sign
to indicate either heavy stained or very faint bands,
the results can be described as follows. From 224
horse samples investigated 218 (97.5 per cent) re-
presented the normal A,A,-type, 5 animals (2.2
per cent) belonged to the A,+A,--type and only
one animal (0.3 per cent) to the A,-&'-type.
The results of mdes revealed that all 52 samples
investfgated represented the A,-A,+-type with proportional staining ability of both bands aImost
identical to the one horse sample mentioned. Samples of 38 donkeys showed only one band which could
be classified a s A,-type, the band migrating slightly
faster tb.n the A,-band of the horse.
In zebras three phenotypes could be established
which are according to staining ability and migration rate the same as the horse haernoglobins. Of
30 zebras samples available for haemoglobin determination 19 (63.3 per cent) showed the normal
A,&-horse type, 10 (33.3 per cent) the k,'A,-horse type and the remaining sample (4.3 per cent)
only one component comparable to the A,-horse
consisted also of two bands and the m x j w bsnd of
the t w was
~ named according to its migratfon speed
in agreflmcnt with thc existing nomenclature
(Braend and Stormont, 19641. A new type having
a double band in the region of t h e m a j x D b m d
in horses appeared rather frequently and i t was
decided to name the allele re2p~nsible for these
two bands TPd, becluse it appeared in homozygous
form only in donkeys and in hetemzygous form
only in mules.
In zebras ts-;z~ new cornbjn3tions of b?n3= werc
additionally found, appnrentIy one allele ( T P z )
being responsible for twa band: migrating faster
than the Dd-bands and one {TfC) being responsible
for two bands with the msjor band migrating between the F- and H-bands of horses.
Examples of migration rate: of t2e new types are
given in Fig. 2 and Fig. 3.
type.
ResuIts of transferrfn determinations are presented
in Table 1.
TABLE 1. Distribution of transferrin types in
equf dae
Horse;
(2301
Donkeys
(35)
Mules
Zebras
(501
(381
Type No.
Type No.
Type No.
Type No.
DD
29
DdDd 19
DaD
8
PF
54
DdH
2
D F
HH
5
Dd1
1
Q0
2
DdM
RR
2
DF
72
DR
11
DO
9
DR
DG
2
12
GG
2
DdI-I
2
GR
3
2
DdO
2
GX
2
DdO
2
DdR
1
GJ
3
DdR
2
DI
1
GM
1
1
DJ
1
GO
l
60
1
DM
6
HH
2
5
HR
2
F1
3
H3
2
FH
16:
10
3
Fd
1
H0
1
F0
l3
FM
9
HR
2
FR
10
FT1
1
I1
2
HP,
3
HI
1
IJ
4
OR
3
HM
1
IM
2
MO
1
JJ
2
JM
1
MO
1
00
1
i3R
1
d
z
Figure 2, New homozggaus transferrin types
I
*
DC
A great variation in transferrin types was found
and an enlargement of the nomenclature used in
horse transferrins was appropriate. Each new type
--
- . .
O r
D:G
D I DH HH GJ
FK
I 0 RR,
Figure 3. Ncw heterozygous transfcrrin types
The frequencies of the different alleles are combined
for all equidae investigated and shov~ndiagrarnatically in Fig. 4.
Albumin typing allowed clear and simple separation
into three bands which were designated A and B
(acc. to Braend, 1964) and C, making provision for
six aIbumin phenotypes. A photograph showing
these phenotypes is present& in Fig. 5, the distri-
PROTEIN POLY3IORPHtSW I N EQUIDAE
HORsES
'
1
DONKEYS
,_,
MULES
J
Figure 4. Freque11~)nT Lran.;lcsrin allclcs in e q u i d n ~
Figure 5 . Albium types in equirla~
bution of diffe~entphenotypes in the four groups
of animals is shown i n Tablc 2.
TABLE 2. Distribution of albumin types in equidae
Species
No.
of
(hybrid h
xnimal;
Alh?!min tlrpes
"A
1 A B 1 BB I CC I *C 1
BC
Horse
Donkey
Mule
Zebra
40
32
0
0
0
21
0
31
5
DISCUSSION AND CONCLUSIONS
Different authors give different ratios of the t w ~
components A, and A, or horse haemoglobin (Studzinski, 1965; Bracnd and Efiemov, 1965). Apparently these ratios may vary in the same animal;
this was aIso revealed by retesting three samples
which in previous tests showed only one band (A,),
but which in r e p a t e d tests showed an additiona1
slow migrating faint band and had to be classified
A, -A,-.
55
The horse possessing the A,-A,+-type was repeatedly blcd and the correct typing confirmed. Chemically investigations should confirm the similarities of this type and thc haemoglobin type found
in all mulc;. Donkeys posecss only the A,-band, a
configuration not yet scen in horses, zebras or
mules, The zebra samples were also tested by
Schmid and the results confirmed (Schmid and
Osterhoff. 1966).
Although not all transferrin types listed in Table
1 could be controlled by family data or breeding
tests, they can be considered as well established. I n
doubtful cases check runs with serum samples labelled with Fe'wwere performed to ensure correct
identification. Furthermore, several horse serum
samples were checked in the Norwegian laboratory
a n d our correct typing was confinned (Braend.
1966). The major bands of the I and J-types are
very near the major H-band of horses, the naming
being in agreement with the existing nomenclature.
The inheritance of the phenotypic Dd-bands could
be clarified in several donkeys and hybrid families.
If the m a j x band of the M-type found in donkeys,
mules and zebras is jn fact the same as the COF
~e;ponf!ing band i n l?orccs could hoxvcvcr not be
represented in our material. The alleles TP, T P
and TfR were present i n all species (hybrids) investigated.
The great variation in the frequency of transferrin
alleles in zebra at the one hand and the concentration of the alleIes TfQand TfFin horses a t the ether,
which has been found i n many studies (see also
Gahne, 1966), could possibly give some i~dication
with regard to the phylogeny of these speries. It
could be reasoned that along tile Iong road of domestication and selcctjon sevet-a1 of the transfersin
genes origihlly present had been diminished or
were lost completely.
The albumin typing gave clear resuIts proving the
observations of Stormont and Suzuki (1963) that
the phenotypes A, AB and B were inherited as if
controlled b y a pair of codominat autosomal alleles.
The extension of that genetic system to three alleles
is appropriate. Frequency calculations revealed similar distributions of AlbA and AlbB in horses and
mules appearing in the latter only in heterozygous
f o r m together with Albc. No difference between
the phenotypes C in donkeys and zebras couId be
estabIished be cause the above M-phenotype was not
between these species. The zebra phenotype AC was
n D t observed, but should be found in a extended
material.
ABSTRACT
Blood and cerurn samples of 234 horses, 40 donkeys,
52 mules and 38 zebras were collected and protein
polymorphism was studied by means of starch gel
dectroplloresf S.
Three haemoglobin phenotypes could be established
in horses, one of them representing a new combination of a major b m d migrating slower than a
secondary faint band. This type was also found in
all mules while a11 donkeys possessed only the
slower migrating band. Tn zebras three types were
found being similar to the phenotypes descrjbed
earlier in horses.
F o u r t e e n transferrin phenotypes were found in horses representing a11 known types except the phenot y p s H 0 end all combinations of the H-type. New
types were discovered in mules, donkeys and zebras
which were named in accordance with the existing
nomenclature for horse transferrjns. The greatest
diversity in transferrin types was found in zebras,
followed by mules combining the transferrin types
of donkeys and horses in heterozygous form. The
gene frequencies of T P and TfF in horses and TP
in donkeys were prevailing.
Albumin studies revealed a clear sepamtion into
three phenotypic bands A, B and C,giving the following phenotypic combinations for the three species and the hybrids: horses AA, AB and BB; donkeys
only CC; rnuIes - AC and BC; zebras BC and CC with AC not yet found.
The usefulness of t!~esestudies in phylogenic fnvestigations is indicated.
-
SUMMARY
Protein polyrnorphlsm In horses, mules, donkeys and
zebras was studied by means of starch gel electrophoreds.
Three haemogIobin phenotypes could be estabIished in
horses, one of them representing a new combinaiion of
a major band migrating slower than a secondary faint
band. This type was also found in all mules, while all
donkeys possessed only the slower mtgrating band. I n
zebras three types were established being similar to the
phenotypes earlier described in horses.
Most of the known transfprrin phenotypes could be
estnblished in horses. New types were discovered in
mules, donkeys and zebras which were named in accordance with the esis:ing nomenclature.
AIbumin studies resulted in the identification of three
bands and n clear species differentiation could be acKeved.
ACKNOWLEDGEMENTS
The Secretary for South West Africa i s thanked
for permitting the pubIlcarion of this paper.
The Director of Nature Conservation and Tourism,
Mr. B. J. G. de 13 Bat and the Assistant Director
af Agriculture, Dr. J. B. Viljoen are thanked for
their co-operation.
Further thanks are due t o Di-. H. Ebedes, Etosha
Game Reserve, South West Africa for kindly supplying most of the mbra blood samples and to Mr.
I. S. Ward-Cox and J. H. van den Berg for their
skilled technical assistance.
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